Fig 1: Induction of GDF15 and an activation of its downstream signaling molecule during the differentiation of hESCs into BAs. (a) Results of microarray analyses regarding TGFB/BMP/GDF family are shown. The vertical and horizontal axes indicate the signal intensity and the time duration after an induction of differentiation, respectively. The cubic polynomial approximation curve is shown on the graph. (b) Western blotting studies for GDF15 protein and beta-tubulin (TUBB) proteins using the differentiating hESCs or human umbilical vein endothelial cells (HUVEC) as a positive control (upper), Western blotting for phosphorylated SMAD2 (pSMAD2), total SMAD2, and TUBB proteins at indicated time points (lower) are shown. (c) qRT-PCR for PAI mRNA. The vertical axis indicates “fold increments” when compared to the data of the samples at 0 h. Data are shown as mean ± SD (n = 3). (d) Western blotting for PAI precursor protein and mature PAI protein. (e) qRT-PCR for GDF15 mRNA. The vertical axis indicates the fold increments compared to the data of the samples at Day 3. Data are shown as mean ± SD (n = 3). (f) Immunostaining studies for GDF15 protein (green) using the terminally differentiated samples at Day 10. Nuclei were counterstained by DAPI (blue).
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